Abstract: Objective To prepare and screen out the anti-EBV ( Epstein-Barr virus) latent membrane protein-1 monoclonal antibodies against Burkitt’s lymphoma (BL). Methods EBV latent membrane protein-1 ( LMP1) transmembrane domain 5 ( TMD5) was synthesized, and was used as the antigen to prepare the anti-TMD5 monoclonal antibodies by hybridoma method. ELISA assay was used to determine the titer of antibodies in mouse ascites. 7-AAD / Annexin V-PE in combination of flow cytometry (FCM) and JC-1 staining in combination of FCM were used to evaluate the apoptotic activity of BL cell line Daudi cells after treatment of monoclonal antibodies. Western blotting assay was used to determine the expression levels of p38-MAPK, IKK, NF-κB. Results After2 rounds of screening, monoclonal antibodies (R2-2-5E-6C and R2-2-8D-3A) were obtained. Both of those could specifically bind to TMD5 peptide, but had no immune response to GAPDH. Compared with these in the NC group, the percentage of Annexin V + 7-AAD + cells after treatment in the R2-2-5E-6C and R2-2-8D-3A groups were increased, the percentage of cells with JC-1fluorescence intensity less than 104 were increased, and the expression levels of intracellular p38-MAPK, IKK and NF-κB were decreased in the Daudi cells (P < 0. 05). Conclusion The obtainedanti-TMD5 monoclonal antibodies R2-2-5E-6C and R2-2-8D-3A could promote BL apoptosis throughthe inhibition of p38-MAPK / IKK / NF-κB pro-survival signaling pathway mediated by LMP1.